Crystalline n--5-chloro-4-(4-chloro-1-methyl-1h-pyrazol-5-yl)-2-thiophenecarboxamide hydrochloride

ABSTRACT

An improved AKT inhibiting compound, crystalline N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamide hydrochloride.

FIELD OF THE INVENTION

This invention relates to an improved protein kinase B (hereinafterPKB/Akt, PKB or AKT) inhibiting compound, the hydrochloride salt ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidein crystalline form. The compound is represented by Structure I:

The compound of this invention is useful as an inhibitor of AKTactivity, and in the treatment of cancer and arthritis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a Powder X-Ray Diffraction (PXRD) pattern ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidein crystalline form.

FIG. 2 depicts a Powder X-Ray Diffraction (PXRD) pattern ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidein crystalline form.

FIG. 3 depicts a Powder X-Ray Diffraction (PXRD) pattern ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidein amorphous form.

DETAILED DESCRIPTION OF THE INVENTION

N-{(1S)-2-Amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamideis a compound which is disclosed and claimed, along withpharmaceutically acceptable salts thereof, as being useful as aninhibitor of AKT activity, particularly in the treatment of cancer andarthritis, in International Application No. PCT/US08/053269, having anInternational filing date of Feb. 7, 2008; International PublicationNumber WO 08/098104 and an International Publication date of Aug. 14,2008 (compound of Example 96). The entire disclosure of which is herebyincorporated by reference. International Application No. PCT/US08/053269discloses the preparation ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride. However the hydrochloride form prepared by the process inExample 96 of International Application No. PCT/US08/053269 isamorphous.

It has now been found that the hydrochloride salt ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidecan be prepared in crystalline form and that the crystalline form hasnumerous advantages over the free acid and other salt forms, includingthe amorphous hydrochloride form. The crystalline form ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride is more readily formulated into pharmaceutical dosageforms, particularly tablets and has improved chemical stability. Thecrystalline form is a non-hygroscopic anhydrous form, and exhibits goodsolubility in bio-relevant media. The amorphous form is hygroscopic.

While the free compound is highly useful as an inhibitor of AKTactivity, particularly in treating cancer and arthritis, the crystallineform of the hydrochloride salt ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehas added advantages.

The compound of this invention,N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride in crystalline form (hereinafter—“Active Ingredient” or“Compound A”), is useful as an inhibitor of AKT activity, particularlyin the treatment of cancer and arthritis. The Active Ingredient can beadministered in a conventional dosage form prepared by combining theActive Ingredient with a conventional pharmaceutically acceptablecarrier or diluent according to techniques readily known to those ofskill in the art, such as those described in International ApplicationNo. PCT/US08/053269.

Suitably, the present invention includes within its scope pharmaceuticalcompositions comprisingN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride in crystalline form, as the Active Ingredient, inassociation with a pharmaceutically acceptable carrier or diluent.Compound A of this invention can be administered by oral, parenteral,intradermal or topical routes of administration. The term parenteral asused herein includes intravenous, intramuscular, subcutaneous,intranasal, intrarectal, intravaginal and intraperitonealadministration. Oral administration is generally preferred. Compound Acan be formulated in dosage forms appropriate for each route ofadministration including capsules, tablets, pills, powders and granules.In such solid dosage forms, the active compound is generally admixedwith at least one inert diluent. The oral dosage forms can alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., lubricating agents, glidants and antioxidants. In thecase of capsules, tablets and pills, the dosage forms may also comprisebuffering agents. Tablets and pills can additionally be prepared for asustained release.

Preparations according to this invention for parenteral administrationinclude sterile aqueous solutions although nonaqueous suspensions ofemulsions can be employed. Such dosage forms may also contain adjuvantssuch as preserving, wetting, osmotic, buffering, emulsifying anddispersing agents. They may be sterilized by, for example, filtrationthrough a bacteria retaining filter, by incorporating sterilizing agentsinto the compositions. Irradiating the compositions or by heating thecompositions.

Doses of the presently invented Active Ingredient in a pharmaceuticaldosage unit as described above will be an efficacious quantitypreferably selected from the range of 0.001-100 mg/kg of total bodyweight, preferably 0.001-50 mg/kg. When treating a human patient in needof AKT inhibition, in need of treatment for cancer, in need of treatmentfor arthritis, the selected dose is preferably administered from 1-6times daily, orally or parenterally. Preferred forms of parenteraladministration include topically, rectally, transdermally, by injectionand continuously by infusion. Oral dosage units for human administrationpreferably contain from 0.05 to 3500 mg of Active Ingredient, mostpreferably from 0.5 to 1,000 mg of Active Ingredient. Oraladministration, which uses lower dosages is preferred. Parenteraladministration, at high dosages, however, also can be used when safe andconvenient for the patient. The above dosages relate to the preferredamount of the Active Ingredient expressed as the free acid.

It will be recognized by one of skill in the art that the optimalquantity and spacing of individual dosages of the Active Ingredient willbe determined by the nature and extent of the condition being treated,the form, route and site of administration, and the particular patientbeing treated, and that such optimums can be determined by conventionaltechniques. It will also be appreciated by one of skill in the art thatthe optimal course of treatment, i.e., the number of doses of the ActiveIngredient given per day for a defined number of days, can beascertained by those skilled in the art using conventional course oftreatment determination tests.

Generally speaking, the compound of this invention is prepared bydissolving the free base,N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamide,in an appropriate organic solvent, such as methyl t-butyl ether (MTBE),then adding to this solution HCl, for example, 4M HCl in dioxane, withheating, for example to 42° C. for 4 hours. The solution is allowed tocool and the compound of this invention is filtered off and dried, forexample, dried in vacuo or air dried at an elevated temperature.

Organic solvents are available from the Aldrich Chemical Company,Milwaukee, Wis.

Because the pharmaceutically active compound of the present invention isactive as an inhibitor or AKT it exhibits therapeutic utility intreating cancer and arthritis.

Suitably, the present invention relates to a method for treating orlessening the severity of a cancer selected from: brain (gliomas),glioblastomas, Bannayan-Zonana syndrome, Cowden disease,Lhermitte-Duclos disease, breast, inflammatory breast cancer, Wilm'stumor, Ewing's sarcoma, Rhabdomyosarcoma, ependymoma, medulloblastoma,colon, head and neck, kidney, lung, liver, melanoma, ovarian,pancreatic, prostate, sarcoma, osteosarcoma, giant cell tumor of bone,thyroid.

Lymphoblastic T cell leukemia, Chronic myelogenous leukemia, Chroniclymphocytic leukemia, Hairy-cell leukemia, acute lymphoblastic leukemia,acute myelogenous leukemia, Chronic neutrophilic leukemia, Acutelymphoblastic T cell leukemia, Plasmacytoma, Immunoblastic large cellleukemia, Mantle cell leukemia, Multiple myeloma Megakaryoblasticleukemia, multiple myeloma, acute megakaryocytic leukemia, promyelocyticleukemia, Erythroleukemia,

malignant lymphoma, hodgkins lymphoma, non-hodgkins lymphoma,lymphoblastic T cell lymphoma, Burkitt's lymphoma, follicular lymphoma,neuroblastoma, bladder cancer, urothelial cancer, lung cancer, vulvalcancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma,esophageal cancer, salivary gland cancer, hepatocellular cancer, gastriccancer, nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST(gastrointestinal stromal tumor) and testicular cancer.

As disclosed in PCT/US2008/053269, the compound of this inventionprepared as in PCT/US2008/053269 was tested for activity generallyaccording to the AKT enzyme assays disclosed in PCT/US2008/053269, andin at least one experimental run exhibited a pIC50 value: equal to 9.0against full length AKT1; equal to 8.0 against full length AKT2; andequal to 8.8 against full length AKT3.

By the term “treating” and derivatives thereof as used herein, is meanttherapeutic therapy. Prophylactic therapy is appropriate, for example,when a subject is considered at high risk for developing cancer, or whena subject has been exposed to a carcinogen.

As used herein, the term “effective amount” and derivatives thereofmeans that amount of a drug or pharmaceutical agent that will elicit thebiological or medical response of a tissue, system, animal or human thatis being sought, for instance, by a researcher or clinician.Furthermore, the term “therapeutically effective amount” and derivativesthereof means any amount which, as compared to a corresponding subjectwho has not received such amount, results in improved treatment,healing, prevention, or amelioration of a disease, disorder, or sideeffect, or a decrease in the rate of advancement of a disease ordisorder. The term also includes within its scope amounts effective toenhance normal physiological function.

By the term “co-administering” and derivatives thereof as used herein ismeant either simultaneous administration or any manner of separatesequential administration of Compound A and a further active ingredientor ingredients, known to be useful in the treatment of cancer, includingchemotherapy and radiation treatment, or to be useful in the treatmentof arthritis. The term further active ingredient or ingredients, as usedherein, includes any compound or therapeutic agent known to or thatdemonstrates advantageous properties when administered to a patient inneed of treatment for cancer or arthritis. Preferably, if theadministration is not simultaneous, the compounds are administered in aclose time proximity to each other. Furthermore, it does not matter ifthe compounds are administered in the same dosage form, e.g. onecompound may be administered topically and another compound may beadministered orally.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be co-administered in the treatmentof cancer in the present invention. Examples of such agents can be foundin Cancer Principles and Practice of Oncology by V. T. Devita and S.Hellman (editors), 6^(th) edition (Feb. 15, 2001), Lippincott Williams &Wilkins Publishers. A person of ordinary skill in the art would be ableto discern which combinations of agents would be useful based on theparticular characteristics of the drugs and the cancer involved. Typicalanti-neoplastic agents useful in the present invention include, but arenot limited to, anti-microtubule agents such as diterpenoids and vincaalkaloids; platinum coordination complexes; alkylating agents such asnitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, andtriazenes; antibiotic agents such as anthracyclins, actinomycins andbleomycins; topoisomerase II inhibitors such as epipodophyllotoxins;antimetabolites such as purine and pyrimidine analogues and anti-folatecompounds; topoisomerase I inhibitors such as camptothecins; hormonesand hormonal analogues; signal transduction pathway inhibitors;non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeuticagents; proapoptotic agents; and cell cycle signaling inhibitors.

Examples of a further active ingredient or ingredients (anti-neoplasticagent) for use in combination or co-administered with the presentlyinvented Compound A are chemotherapeutic agents.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem. Soc., 93:2325. 1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl. Acad. Sci. USA,77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar,J. Biol. Chem. 256: 10435-10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LaQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaleJournal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann.Intern. Med., 111:273, 1989) and for the treatment of breast cancer(Holmes et al., J. Nat. Cancer Inst., 83:1797, 1991.) It is a potentialcandidate for treatment of neoplasms in the skin (Einzig et. al., Proc.Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastireet. al., Sem. Oncol., 20:56, 1990). The compound also shows potentialfor the treatment of polycystic kidney disease (Woo et al., Nature,368:750, 1994), lung cancer and malaria. Treatment of patients withpaclitaxel results in bone marrow suppression (multiple cell lineages,Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide, 1998) related tothe duration of dosing above a threshold concentration (50 nM) (Kearns,C. M. et. al., Seminars in Oncology. 3(6) p. 16-23, 1995).

Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine, N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkle plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine, 3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)], commerciallyavailable as an injectable solution of vinorelbine tartrate(NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine isindicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo, aquation and form intra- and interstrandcrosslinks with DNA causing adverse biological effects to the tumor.Examples of platinum coordination complexes include, but are not limitedto, cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′], is commercially available asPARAPLATIN® as an injectable solution. Carboplatin is primarilyindicated in the first and second line treatment of advanced ovariancarcinoma. Bone marrow suppression is the dose limiting toxicity ofcarboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-resectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also know as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins.

Epipodophyllotoxins are phase specific anti-neoplastic agents derivedfrom the mandrake plant. Epipodophyllotoxins typically affect cells inthe S and G₂ phases of the cell cycle by forming a ternary complex withtopoisomerase II and DNA causing DNA strand breaks. The strand breaksaccumulate and cell death follows. Examples of epipodophyllotoxinsinclude, but are not limited to, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leucopenia tends to be more severethan thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leucopenia and thrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mercaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-β-D-arabinofuranosyl-2(1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leucopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of thioguanine administration.However, gastrointestinal side effects occur and can be dose limiting.Other purine analogs include pentostatin, erythrohydroxynonyladenine,fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of gemcitabine administration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leucopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl, (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxyl]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®.

Irinotecan is a derivative of camptothecin which binds, along with itsactive metabolite SN-38, to the topoisomerase I—DNA complex. It isbelieved that cytotoxicity occurs as a result of irreparable doublestrand breaks caused by interaction of the topoisomerase I: DNA:irintecan or SN-38 ternary complex with replication enzymes. Irinotecanis indicated for treatment of metastatic cancer of the colon or rectum.The dose limiting side effects of irinotecan HCl are myelosuppression,including neutropenia, and GI effects, including diarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as theinjectable solution HYCAMTIN®. Topotecan is a derivative of camptothecinwhich binds to the topoisomerase I—DNA complex and prevents relegationof singles strand breaks caused by Topoisomerase I in response totorsional strain of the DNA molecule. Topotecan is indicated for secondline treatment of metastatic carcinoma of the ovary and small cell lungcancer. The dose limiting side effect of topotecan HCl ismyelosuppression, primarily neutropenia.

Also of interest, is the camptothecin derivative of formula A following,currently under development, including the racemic mixture (R,S) form aswell as the R and S enantiomers:

known by the chemical name″7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R,S)-camptothecin(racemic mixture) or″7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(R)-camptothecin(R enantiomer) or″7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20(S)-camptothecin(S enantiomer). Such compound as well as related compounds aredescribed, including methods of making, in U.S. Pat. Nos. 6,063,923;5,342.947; 5,559,235; 5,491,237 and pending U.S. patent application Ser.No. 08/977,217 filed Nov. 24, 1997.

Hormones and hormonal analogues are useful compounds for treatingcancers in which there is a relationship between the hormone(s) andgrowth and/or lack of growth of the cancer. Examples of hormones andhormonal analogues useful in cancer treatment include, but are notlimited to, adrenocorticosteroids such as prednisone and prednisolonewhich are useful in the treatment of malignant lymphoma and acuteleukemia in children; aminoglutethimide and other aromatase inhibitorssuch as anastrozole, letrazole, vorazole, and exemestane useful in thetreatment of adrenocortical carcinoma and hormone dependent breastcarcinoma containing estrogen receptors; progestrins such as megestrolacetate useful in the treatment of hormone dependent breast cancer andendometrial carcinoma; estrogens, androgens, and anti-androgens such asflutamide, nilutamide, bicalutamide, cyproterone acetate and5α-reductases such as finasteride and dutasteride, useful in thetreatment of prostatic carcinoma and benign prostatic hypertrophy;anti-estrogens such as tamoxifen, toremifene, raloxifene, droloxifene,iodoxyfene, as well as selective estrogen receptor modulators (SERMS)such those described in U.S. Pat. Nos. 5,681,835, 5,877,219, and6,207,716, useful in the treatment of hormone dependent breast carcinomaand other susceptible cancers; and gonadotropin-releasing hormone (GnRH)and analogues thereof which stimulate the release of leutinizing hormone(LH) and/or follicle stimulating hormone (FSH) for the treatmentprostatic carcinoma, for instance, LHRH agonists and antagonists such asgoserelin acetate and luprolide.

Signal transduction pathway inhibitors are those inhibitors, which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation. Signaltransduction inhibitors useful in the present invention includeinhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases,SH2/SH3 domain blockers, serine/threonine kinases, phosphatidylinositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

Several protein tyrosine kinases catalyse the phosphorylation ofspecific tyrosyl residues in various proteins involved in the regulationof cell growth. Such protein tyrosine kinases can be broadly classifiedas receptor or non-receptor kinases.

Receptor tyrosine kinases are transmembrane proteins having anextracellular ligand binding domain, a transmembrane domain, and atyrosine kinase domain. Receptor tyrosine kinases are involved in theregulation of cell growth and are generally termed growth factorreceptors. Inappropriate or uncontrolled activation of many of thesekinases, i.e. aberrant kinase growth factor receptor activity, forexample by over-expression or mutation, has been shown to result inuncontrolled cell growth. Accordingly, the aberrant activity of suchkinases has been linked to malignant tissue growth. Consequently,inhibitors of such kinases could provide cancer treatment methods.Growth factor receptors include, for example, epidermal growth factorreceptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2,erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosinekinase with immunoglobulin-like and epidermal growth factor homologydomains (TIE-2), insulin growth factor-I (IGFI) receptor, macrophagecolony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growthfactor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin(eph) receptors, and the RET protooncogene. Several inhibitors of growthreceptors are under development and include ligand antagonists,antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.Growth factor receptors and agents that inhibit growth factor receptorfunction are described, for instance, in Kath, John C., Exp. Opin. Ther.Patents (2000) 10(6):803-818; Shawver et al DDT Vol 2, No. 2 Feb. 1997;and Lofts. F. J. et al, “Growth factor receptors as targets”, NewMolecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr,David, CRC press 1994, London.

Tyrosine kinases, which are not growth factor receptor kinases aretermed non-receptor tyrosine kinases. Non-receptor tyrosine kinases foruse in the present invention, which are targets or potential targets ofanti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focaladhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Suchnon-receptor kinases and agents which inhibit non-receptor tyrosinekinase function are described in Sinh. S. and Corey, S. J., (1999)Journal of Hematotherapy and Stem Cell Research 8 (5): 465-80; andBolen, J. B., Brugge, J. S., (1997) Annual review of Immunology. 15:371-404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domainbinding in a variety of enzymes or adaptor proteins including, PI3-K p85subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2) andRas-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussedin Smithgall, T. E. (1995), Journal of Pharmacological and ToxicologicalMethods. 34(3) 125-32.

Inhibitors of Serine/Threonine Kinases including MAP kinase cascadeblockers which include blockers of Raf kinases (rafk), Mitogen orExtracellular Regulated Kinase (MEKs), and Extracellular RegulatedKinases (ERKs); and Protein kinase C family member blockers includingblockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).IkB kinase family (IKKa, IKKb), PKB family kinases, akt kinase familymembers, and TGF beta receptor kinases. Such Serine/Threonine kinasesand inhibitors thereof are described in Yamamoto. T., Taya, S.,Kalbuchi, K., (1999), Journal of Biochemistry. 126 (5) 799-803; Brodt.P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60,1101-1107; Massague, J., Weis-Garcia. F. (1996) Cancer Surveys.27:41-64; Philip, P. A., and Harris, A. L. (1995), Cancer Treatment andResearch. 78: 3-27, Lackey, K. et al Bioorganic and Medicinal ChemistryLetters. (10), 2000, 223-226; U.S. Pat. No. 6,268,391; andMartinez-Iacaci, L., et al, Int. J. Cancer (2000). 88(1), 44-52.

Inhibitors of Phosphotidyl inositol-3 Kinase family members includingblockers of PI3-kinase, ATM, DNA-PK, and Ku may also be useful in thepresent invention. Such kinases are discussed in Abraham, R. T. (1996),Current Opinion in Immunology. 8 (3) 412-8; Canman, C. E., Lim, D. S.(1998), Oncogene 17 (25) 3301-3308; Jackson. S. P. (1997), InternationalJournal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. etal, Cancer res, (2000) 60(6), 1541-1545.

Also of interest in the present invention are Myo-inositol signalinginhibitors such as phospholipase C blockers and Myoinositol analogues.Such signal inhibitors are described in Powis, G., and Kozikowski A.,(1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workmanand David Kerr, CRC press 1994. London.

Another group of signal transduction pathway inhibitors are inhibitorsof Ras Oncogene. Such inhibitors include inhibitors offarnesyltransferase, geranyl-geranyl transferase, and CAAX proteases aswell as anti-sense oligonucleotides, ribozymes and immunotherapy. Suchinhibitors have been shown to block ras activation in cells containingwild type mutant ras, thereby acting as antiproliferation agents. Rasoncogene inhibition is discussed in Scharoveky, O. G., Rozados, V. R.,Gervasoni, S. I. Mater, P. (2000). Journal of Biomedical Science. 7(4)292-8; Ashby. M. N. (1998). Current Opinion in Lipidology. 9 (2) 99-102;and BioChim. Biophys. Acta, (19899) 1423(3):19-30.

As mentioned above, antibody antagonists to receptor kinase ligandbinding may also serve as signal transduction inhibitors. This group ofsignal transduction pathway inhibitors includes the use of humanizedantibodies to the extracellular ligand binding domain of receptortyrosine kinases. For example Imclone C225 EGFR specific antibody (seeGreen, M. C. et al, Monoclonal Antibody Therapy for Solid Tumors, CancerTreat. Rev., (2000), 26(4), 269-286); Herceptin® erbB2 antibody (seeTyrosine Kinase Signalling in Breast cancer:erbB Family ReceptorTyrosine Kinases, Breast Cancer Res., 2000, 2(3), 176-183); and 2CBVEGFR2 specific antibody (see Brekken, R. A. et al, Selective Inhibitionof VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumorgrowth in mice, Cancer Res. (2000) 60, 5117-5124).

Non-receptor kinase angiogenesis inhibitors may also be useful in thepresent invention. Inhibitors of angiogenesis related VEGFR and TIE2 arediscussed above in regard to signal transduction inhibitors (bothreceptors are receptor tyrosine kinases). Angiogenesis in general islinked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR havebeen shown to inhibit angiogenesis, primarily VEGF expression.Accordingly, non-receptor tyrosine kinase inhibitors may be used incombination with the compounds of the present invention. For example,anti-VEGF antibodies, which do not recognize VEGFR (the receptortyrosine kinase), but bind to the ligand; small molecule inhibitors ofintegrin (alpha, beta₃) that will inhibit angiogenesis; endostatin andangiostatin (non-RTK) may also prove useful in combination with thedisclosed compounds. (See Bruns C J et al (2000), Cancer Res., 60:2926-2935; Schreiber A B. Winkler M E, and Derynck R. (1986), Science,232: 1250-1253; Yen L et al. (2000). Oncogene 19: 3460-3469).

Agents used in immunotherapeutic regimens may also be useful in thepresent invention. There are a number of immunologic strategies togenerate an immune response. These strategies are generally in the realmof tumor vaccinations. The efficacy of immunologic approaches may begreatly enhanced through combined inhibition of signaling pathways usinga small molecule inhibitor. Discussion of the immunologic/tumor vaccineapproach against erbB2/EGFR are found in Reilly R T et al. (2000),Cancer Res. 60: 3569-3576; and Chen Y, Hu D, Eling D J, Robbins J, andKipps T J. (1998). Cancer Res. 58: 1965-1971.

Agents used in proapoptotic regimens (e.g., bcl-2 antisenseoligonucleotides) may also be used in the combination of the presentinvention. Members of the Bcl-2 family of proteins block apoptosis.Upregulation of bcl-2 has therefore been linked to chemoresistance.Studies have shown that the epidermal growth factor (EGF) stimulatesanti-apoptotic members of the bcl-2 family (i.e., mcl-1). Therefore,strategies designed to downregulate the expression of bcl-2 in tumorshave demonstrated clinical benefit and are now in Phase II/III trials,namely Genta's G3139 bcl-2 antisense oligonucleotide. Such proapoptoticstrategies using the antisense oligonucleotide strategy for bcl-2 arediscussed in Water J S et al. (2000). J. Clin. Oncol. 18: 1812-1823; andKitada S et al. (1994), Antisense Res. Dev. 4: 71-79.

Cell cycle signalling inhibitors inhibit molecules involved in thecontrol of the cell cycle. A family of protein kinases called cyclindependent kinases (CDKs) and their interaction with a family of proteinstermed cyclins controls progression through the eukaryotic cell cycle.The coordinate activation and inactivation of different cyclin/CDKcomplexes is necessary for normal progression through the cell cycle.Several inhibitors of cell cycle signalling are under development. Forinstance, examples of cyclin dependent kinases, including CDK2, CDK4,and CDK6 and inhibitors for the same are described in, for instance,Rosania et al, Exp. Opin. Ther. Patents (2000) 10(2):215-230.

In one embodiment, the cancer treatment method of the claimed inventionincludes the co-administration of Compound A and at least oneanti-neoplastic agent, such as one selected from the group consisting ofanti-microtubule agents, platinum coordination complexes, alkylatingagents, antibiotic agents, topoisomerase II inhibitors, antimetabolites,topoisomerase I inhibitors, hormones and hormonal analogues, signaltransduction pathway inhibitors, non-receptor tyrosine kinaseangiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents,and cell cycle signaling inhibitors.

Compound A of this invention is useful as an Inhibitor of AKT, in thetreatment of cancer and arthritis in mammals, particularly humans, inneed thereof.

The method of this invention of treating cancer and arthritis inmammals, including humans, comprises administering to a subject in needof such treatment a therapeutically effective amount of Compound A ofthe present invention.

The invention also provides for the use of Compound A in the manufactureof a medicament for use in therapy.

The invention also provides for the use of Compound A in the manufactureof a medicament for use as an AKT inhibitor.

The invention also provides for the use of Compound A in the manufactureof a medicament for use in treating cancer.

The invention also provides for the use of Compound A in the manufactureof a medicament for use in treating arthritis.

The invention also provides for a pharmaceutical composition for use inthe treatment of cancer which comprises Compound A and apharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use inthe treatment of arthritis which comprises Compound A and apharmaceutically acceptable carrier.

The following examples further illustrate the present invention. Theexamples are not intended to limit the scope of the invention as definedhereinabove and as claimed below.

Example 1 Preparation of: CrystallineN-{(1S)-2-Amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride

AmorphousN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidefree base is prepared as described in International Application No.PCT/US08/053269. 1 mL of methyl t-butyl ether (MTBE) is added to a vialcontaining 50.8 mg of amorphousN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidefree base. Most of the solid dissolved. After addition of the HCl acid(1 eq., 29.7 mL of 4M HCl in 1,4-dioxane) to the reaction mixture, awhite solid material crashed out quickly at room temperature. An aliquotof the reaction mixture was analyzed by polarized light microscopy (PLM)and the solid was found to be amorphous. The slurry was then heated to40° C. with magnetic stirring to dissolve most of the solid material.The thin slurry was stirred at 40° C. overnight, and then cooled from40° C. to 25° C. A thick white slurry formed. An aliquot of the reactionmixture was analyzed by polarized light microscopy (PLM) and the solidwas found to be crystalline. The white solid was collected by vacuumfiltration, and then dried at 50° C. overnight. The product wasdetermined be a crystalline solid by the Powder X-Ray Diffraction (PXRD)pattern depicted in FIG. 1.

Instrument: Rigaku Miniflex II, Serial DD02652 Key Operating Parameters:Method Normal Run

The sample is scanned using the following parameters:Scan range: 2-40 degrees two-thetaGenerator power: 30 kV, 15 mA

Radiation Source: Cu KAlpha

Scan mode: FTCount time: 1.0 secondsStep width: 0.020 degrees two-theta per stepScan axis: 2theta/thetaIncident Beam optics: ±2.5° divergence angle soller slit, 1.25 degreedivergent slit, 0.3 mmRecSlit, 1.25 degrees anti-scatter slitDiffracted Beam optics: fixed slits (MiniFlex II), ±2.5° soller slits

Detector Type: Rigaku MiniFlex II SC (Scintillator Counter)

The sample is packed into a zero background sample holder (100 μmindent) and gently flattened using a glass slide.

Example 2 Preparation of: CrystallineN-{(1S)-2-Amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride

5 mL of MTBE was added to 213.9 mg ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamideamorphous free-base (0.500 mMol.). The mixture was heated to 40° C. withmagnetic stirring for 1 hour. A solution of 4M HCl in 1,4-dioxane (1 eq;125.1 uL) was added in four equal portions. After addition of the firstportion (0.25 eq), the mixture was heated to 42° C. and 1 mL ofacetonitrile was added to dissolve all of the solid material. After theremainder of the HCl solution was added, some solid material appeared.The slurry was stirred at 42° C. for 4 hours, then was cooled slowly to22° C. overnight with 90 minute holds at 35° C., 30° C., and 25° C. Thewhite solid material was filtered and dried at 50° C. under vacuum witha slow nitrogen bleed overnight. The yield was 73.6% (0.3685 mmol;170.90 mg) of the HCl salt. The solid was found to be 1:1 stoichiometricHCl salt by ion chromatography, and to be crystalline by the PowderX-Ray Diffraction (PXRD) pattern depicted in FIG. 2 and as characterizedby diffraction peaks below, and to have a melting point of 211° C.

PXRD Peaks (Values Given in Degrees Two-Theta with d-Spacing inParenthesis):

 7.2(12.20) 14.4(6.16) 17.9(4.94) 18.5(4.79) 20.8(4.26) 21.5(4.12)22.4(3.96) 22.9(3.88) 23.7(3.75) 24.5(3.63) 24.7(3.61) 25.1(4.12)25.7(3.46) 27.3(3.26) 28.2(3.16) 28.8(3.10) 30.4(2.94) 32.4(2.76)32.7(2.73) 35.2(2.55) 36.1(2.48) 40.0(2.25) 41.3(2.18) 41.7(2.16)Instrument: PANalytical X'Pert-Pro MPD with Johansson Kα1 monochromator,using X'Celerator detector

Key Operating Parameters:

Radiation: Cu (Kα1), 1.540598 angstroms (monochromatic)

Detector: X'Celerator Tension: 45 kV Current: 40 mA

Start angle: 2.00° 2θEnd angle: 52.0° 2θStep size: 0.02°

Time/step: 40.0 sec

Scan speed: 0.05°/secIncident beam: 2° fixed anti-scatter slit, and programmable divergenceslitDiffracted beam: 0.02 rad soller slit, and programmable anti-scatterslitSamples prepared on silicon zero background sample holder.

Example 3 Preparation of AmorphousN{(1S)-2-Amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride

The hydrochloride salt ofN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamideprepared generally as described in Example 96 of InternationalApplication No. PCT/US2008/053269, having an International Filing dateof Feb. 7, 2008, and having International Publication NumberWO/2008/098104 and an International Publication Date of Aug. 14, 2008,which generally states that the free base compound,N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamide,is treated with 4M HCl in dioxane and after 5 min, the solution isconcentrated and dried under vacuum to affordN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride, yielded an amorphous solid as indicated by the PowderX-Ray Diffraction (PXRD) pattern depicted in FIG. 3.

Instrument: Rigaku Miniflex II, Serial DD02652 Key Operating Parameters:Method Normal Run

The sample is scanned using the following parameters:Scan range: 2-40 degrees two-thetaGenerator power: 30 kV, 15 mA

Radiation Source: Cu KAlpha

Scan mode: FTCount time: 1.0 secondsStep width: 0.020 degrees two-theta per stepScan axis: 2theta/thetaIncident Beam optics: ±2.5° divergence angle soller slit, 1.25 degreedivergent slit, 0.3 mmRecSlit, 1.25 degrees anti-scatter slitDiffracted Beam optics: fixed slits (MiniFlex II), ±2.5° soller slits

Detector Type: Rigaku MiniFlex II SC (Scintillator Counter)

The sample is packed into a glass sample holder and gently flattenedusing a glass slide.

Example 4 Tablet Composition

Lactose, microcrystalline cellulose, sodium starch glycolate, magnesiumstearate and crystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methy-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride are blended in the proportions shown in Table 1 below. Theblend is then compressed into tablets.

TABLE 1 INGREDIENT mg. crystalline N-{(1S)-2-amino-1-[(3- 8.45fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2- thiophenecarboxamide hydrochloridemicrocrystalline cellulose 112 lactose 70 sodium starch glycolate 8magnesium stearate 2

Example 5 Injectable Parenteral Composition

An injectable form for administering the active Ingredient is producedby stirring 5.0 mg. of crystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride in 1.0 ml. of normal saline.

Example 6 Hygroscopicity Crystalline

CrystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride made generally according to Example 1, and confirmed to becrystalline by PXRD, was tested using a VTI instrument SGA 100 using thefollowing method.

Sample Preparation: Solvent: Water

Drying Temp (° C.): No drying

Experiment Temp (° C.): 25

Max Equilibrium Time (min) 240

Equilibrium Criteria (wt %): 0.0025 in 10 min

Data Logging Interval (min): 2 or 0.01 wt %

There were 24 Relative Humidity steps used. The Relative Humidity ateach step is as follows.

1-40; 2-50; 3-60; 4-70; 5-80; 6-90; 7-80; 8-70; 9-60; 10-50;11-40; 12-30; 13-20; 14-10; 15-5; 16-10; 17-20; 18-30; 19-40;20-50; 21-60; 22-70; 23-80; and 24-90.

The weight increase of the compound was less than 1% w/w moisture,indicating the compound has a low level of hygroscopicity.

Amorphous

In a separate experiment on a different day, amorphousN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride (confirmed to be amorphous by PXRD), was tested using aVTI instrument SGA 100 using the following method.

Sample Preparation: Solvent: Water Drying Temp (° C.): 60

Heating Rate (° C./min): 1Max Drying Time (min): 120

Experiment Temp (° C.): 25

Max Equilibrium Time (min) 300

Equilibrium Criteria (wt %): 0.0025 in 10 min

Data Logging Interval (min): 2 or 0.01 wt %

There were 19 Relative Humidity steps used. The Relative Humidity ateach step is as follows.

1-0; 2-10; 3-20; 4-30; 5-40; 6-50; 7-60; 8-70; 9-80; 10-90;11-80; 12-70; 13-60; 14-50; 15-40; 16-30; 17-20; 18-10; and 19-0.

The weight increase of the compound was more than 18% w/w moisture,indicating the compound has a high level of hygroscopicity.

While the preferred embodiments of the invention are illustrated by theabove, it is to be understood that the invention is not limited to theprecise instructions herein disclosed and that the right to allmodifications coming within the scope of the following claims isreserved.

What is claimed is: 1.N-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride in crystalline form.
 2. A pharmaceutical compositioncomprising a compound of claim 1 and a pharmaceutically acceptablecarrier or diluent.
 3. A process for preparing a pharmaceuticalcomposition containing a pharmaceutically acceptable carrier and aneffective amount of a compound of claim 1, which process comprisesbringing the compound of claim 1 into association with apharmaceutically acceptable carrier.
 4. A method of treating orlessening the severity of cancer in a mammal in need thereof, whichcomprises administering to such mammal a therapeutically effectiveamount of a compound of claim
 1. 5. The method of claim 4 wherein themammal is a human.
 6. The method according to claim 5 wherein saidcancer is selected from: brain (gliomas), glioblastomas, Bannayan-Zonanasyndrome, Cowden disease, Lhermitte-Duclos disease, breast, inflammatorybreast cancer, Wilm's tumor, Ewing's sarcoma, Rhabdomyosarcoma,ependymoma, medulloblastoma, colon, head and neck, kidney, lung, liver,melanoma, ovarian, pancreatic, prostate, sarcoma, osteosarcoma, giantcell tumor of bone, thyroid, Lymphoblastic T cell leukemia, Chronicmyelogenous leukemia, Chronic lymphocytic leukemia, Hairy-cell leukemia,acute lymphoblastic leukemia, acute myelogenous leukemia, Chronicneutrophilic leukemia, Acute lymphoblastic T cell leukemia,Plasmacytoma, Immunoblastic large cell leukemia, Mantle cell leukemia,Multiple myeloma Megakaryoblastic leukemia, multiple myeloma, acutemegakaryocytic leukemia, promyelocytic leukemia, Erythroleukemia,malignant lymphoma, hodgkins lymphoma, non-hodgkins lymphoma,lymphoblastic T cell lymphoma, Burkitt's lymphoma, follicular lymphoma,neuroblastoma, bladder cancer, urothelial cancer, lung cancer, vulvalcancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma,esophageal cancer, salivary gland cancer, hepatocellular cancer, gastriccancer, nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST(gastrointestinal stromal tumor) and testicular cancer.
 7. A method oftreating cancer in a mammal in need thereof, which comprises:administering to such mammal a therapeutically effective amount of a) acompound of claim 1; and b) at least one anti-neoplastic agent.
 8. Themethod claim 7, wherein the at least one anti-neoplastic agent isselected from the group consisting essentially of anti-microtubuleagents, platinum coordination complexes, alkylating agents, antibioticagents, topoisomerase II inhibitors, antimetabolites, topoisomerase Iinhibitors, hormones and hormonal analogues, signal transduction pathwayinhibitors; non-receptor tyrosine kinase angiogenesis inhibitors;immunotherapeutic agents; proapoptotic agents; and cell cycle signalinginhibitors.
 9. The method of claim 5 wherein the compound isadministered orally.
 10. A method of treating or lessening the severityof cancer in a mammal in need thereof, which comprises administering tosuch mammal a therapeutically effective amount of a composition of claim2.
 11. The method of claim 10 wherein the mammal is a human.
 12. Themethod according to claim 11 wherein said cancer is selected from: brain(gliomas), glioblastomas, Bannayan-Zonana syndrome, Cowden disease,Lhermitte-Duclos disease, breast, inflammatory breast cancer, Wilm'stumor, Ewing's sarcoma, Rhabdomyosarcoma, ependymoma, medulloblastoma,colon, head and neck, kidney, lung, liver, melanoma, ovarian,pancreatic, prostate, sarcoma, osteosarcoma, giant cell tumor of bone,thyroid, Lymphoblastic T cell leukemia, Chronic myelogenous leukemia,Chronic lymphocytic leukemia, Hairy-cell leukemia, acute lymphoblasticleukemia, acute myelogenous leukemia, Chronic neutrophilic leukemia,Acute lymphoblastic T cell leukemia, Plasmacytoma, Immunoblastic largecell leukemia, Mantle cell leukemia, Multiple myeloma Megakaryoblasticleukemia, multiple myeloma, acute megakaryocytic leukemia, promyelocyticleukemia, Erythroleukemia, malignant lymphoma, hodgkins lymphoma,non-hodgkins lymphoma, lymphoblastic T cell lymphoma, Burkitt'slymphoma, follicular lymphoma, neuroblastoma, bladder cancer, urothelialcancer, lung cancer, vulval cancer, cervical cancer, endometrial cancer,renal cancer, mesothelioma, esophageal cancer, salivary gland cancer,hepatocellular cancer, gastric cancer, nasopharangeal cancer, buccalcancer, cancer of the mouth, GIST (gastrointestinal stromal tumor) andtesticular cancer.
 13. CrystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride of claim 1 having characteristic diffraction peaks at14.4°±0.3° and 32.4°±0.3° in an Powder X-Ray Diffractogram using Cu Kαradiation.
 14. CrystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride having the characteristic diffraction peaks recited inclaim 13 and having characteristic diffraction peaks at 25.1°±0.3° and25.7°±0.3° in the Powder X-Ray Diffractogram using Cu Kα radiation. 15.CrystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride having the characteristic diffraction peaks recited inclaim 14 and having characteristic diffraction peaks at 21.5°±0.3° and20.8°±0.3° in the Powder X-Ray Diffractogram using Cu Kα radiation. 16.A pharmaceutical composition comprising crystallineN-{(1S)-2-amino-1-[(3-fluorophenyl)methyl]ethyl}-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)-2-thiophenecarboxamidehydrochloride having the characteristic diffraction peaks recited inclaim 13 and a pharmaceutically acceptable carrier or diluent.
 17. Amethod of treating or lessening the severity of cancer in a mammal inneed thereof, which comprises administering to such mammal atherapeutically effective amount of a composition of claim
 16. 18. Themethod of claim 17 wherein the mammal is a human.
 19. The method ofclaim 18 wherein the compound is administered orally.
 20. The methodaccording to claim 18 wherein said cancer is selected from: brain(gliomas), glioblastomas, Bannayan-Zonana syndrome, Cowden disease,Lhermitte-Duclos disease, breast, inflammatory breast cancer, Wilm'stumor, Ewing's sarcoma, Rhabdomyosarcoma, ependymoma, medulloblastoma,colon, head and neck, kidney, lung, liver, melanoma, ovarian,pancreatic, prostate, sarcoma, osteosarcoma, giant cell tumor of bone,thyroid, Lymphoblastic T cell leukemia, Chronic myelogenous leukemia,Chronic lymphocytic leukemia, Hairy-cell leukemia, acute lymphoblasticleukemia, acute myelogenous leukemia, Chronic neutrophilic leukemia,Acute lymphoblastic T cell leukemia, Plasmacytoma, Immunoblastic largecell leukemia, Mantle cell leukemia, Multiple myeloma Megakaryoblasticleukemia, multiple myeloma, acute megakaryocytic leukemia, promyelocyticleukemia, Erythroleukemia, malignant lymphoma, hodgkins lymphoma,non-hodgkins lymphoma, lymphoblastic T cell lymphoma, Burkitt'slymphoma, follicular lymphoma, neuroblastoma, bladder cancer, urothelialcancer, lung cancer, vulval cancer, cervical cancer, endometrial cancer,renal cancer, mesothelioma, esophageal cancer, salivary gland cancer,hepatocellular cancer, gastric cancer, nasopharangeal cancer, buccalcancer, cancer of the mouth, GIST (gastrointestinal stromal tumor) andtesticular cancer.